Biosafety Level 2 Standard Safety Requirements
Biosafety in Microbiological and Biomedical Laboratories, (BMBL), Centers for Disease Control and Prevention/National Institutes of Health, 5th edition, U.S. Department of Public Health and Human Services, Dec 2009. Available online at: http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.
NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH Guidelines), April 2002. Available online at: http://oba.od.nih.gov/oba/rac/guidelines_02/NIH_Gdlnes_lnk_2002z.pdf.
Handling and Disposal of Laboratory Sharps, Biosafety Facts, Division of Research Safety, University of Illinois at Urbana-Champaign. Available online at: http://www.drs.illinois.edu/bss/factsheets/.
Treatment and Disposal of Biological Materials, Biosafety Facts, Division of Research Safety, University of Illinois at Urbana-Champaign. Available online at: http://www.drs.illinois.edu/bss/factsheets/.
All project personnel, including students, must be trained and proficient in Biosafety Level 2 practices. A copy of the CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories, 5th edition (BMBL) must be available to all laboratory staff. All persons working in the laboratory areas must be advised of the hazards and nature of the research being conducted. Project approval information and all project protocols must be made available to all project personnel and to laboratory directors in shared or common facilities.
University policy requires that significant research-related incidents be reported immediately to the Institutional Biosafety Committee via the Biological Safety Section (BSS) of the Division of Research Safety. Such incidents include research-related accidents and illnesses as well as inadvertent release or improper disposal of biohazardous or recombinant DNA materials. BSS can be contacted at 333-2755 and via e-mail.
The laboratory director must restrict access to the laboratory and the animal rooms, when work with the Biosafety Level 2 materials is occurring. Access to areas where biological agents or potentially biohazardous materials are stored must be controlled. This may include keeping the laboratory doors locked when laboratory personnel are not in the immediate area and/or maintaining a lock on a freezer storing biohazardous materials that is located in a common-use room or area.
The laboratory director must ensure that all participants, including animal care personnel, and observers of the research procedures wear appropriate protective equipment and follow appropriate safety practices.
Signage and Labels
Biohazard access signs must be posted at the entrances to laboratory areas and animal care rooms where the Biosafety Level 2 materials are used. The Biological Safety Section, Division of Research Safety will provide appropriate access signs. Biohazard stickers should be posted on all equipment used to store or manipulate the Biosafety Level 2 materials. Stickers are available from the Biological Safety Section, 333-2755 or e-mail via e-mail.
All microwaves and refrigerators utilized for laboratory materials should be labeled: “For research purposes only. No food or drink allowed in this refrigerator (or microwave).”
Transportation of Biohazards
If biohazardous materials are transported by laboratory personnel, they must be contained in such a way as to prevent release to the environment in case of an accident (e.g., placed in a primary container that is a securely closed, a watertight tube, vial or ampoule, which is then placed in an unbreakable, watertight secondary container that is labeled with the international biohazard symbol. Enough absorbent material, such as paper towels, must be placed in the space at the top, bottom, and sides between the primary and secondary containers to absorb the entire contents of the primary container in case of breakage or leakage). For intra-facility transport, use of a cart to facilitate transport is recommended. Appropriate personal protective equipment (PPE) and clean-up materials should be kept in the transport vehicle or on the cart used to transport the material.
If a spill occurs during transport, do not attempt to clean it up without appropriate spill response material and PPE. Keep other persons clear of the spill.
Decontamination – work surfaces, equipment and spills
Lab equipment and work surfaces should be decontaminated with an effective disinfectant such as 10% fresh bleach solution with a contact time of at least 10 minutes or with an EPA approved disinfectant. Lab equipment and work surfaces should be decontaminated after work with infectious materials is finished, and especially after overt spills, splashes, or other contamination by infectious materials.
To clean up and decontaminate spills, allow aerosols to settle; wear vinyl, latex, or rubber utility gloves, eye protection and protective clothing; gently cover spill with paper toweling and apply a suitable disinfectant starting at perimeter and working toward the center. Allow an appropriate contact time before clean up. Clean up materials with fresh paper towels. The area of the spill should be re-wiped/mopped with disinfectant after all visible material is removed to ensure complete decontamination. The clean-up materials may be placed in the normal trash.
Cloth material is not appropriate in laboratories because proper decontamination following a spill is very difficult to achieve. It is recommended that any cloth chairs are covered with a durable, leak-resistant material while laboratory work with potentially infectious materials is occurring around them.
Procedures with a potential for creating infectious aerosols or splashes must be performed in a biological safety cabinet (BSC). These may include centrifuging, grinding, blending, vigorous shaking or mixing, sonic disruption, opening containers of infectious materials whose internal pressures may be different from ambient pressures. If a biological safety cabinet is to be used in this project, it must be certified at the beginning of the project, and it must undergo re-certification on an annual basis. Call (333-2755) or e-mail (via email) for information on how to set-up an appointment for certification.
Face protection (goggles and mask, faceshield, and/or other splatter guards) must be provided and used to protect against splashes or sprays of infectious or other hazardous materials to the face, when the biohazardous materials are manipulated outside the BSC.
Protective lab coats should be worn by personnel while in the laboratory. This protective clothing should be removed and left in the laboratory before leaving for non-lab areas (e.g., cafeteria, library, administrative offices). All protective clothing is either disposed of by the lab or laundered – it should never be taken home by personnel.
Gloves must be worn when hands may contact potentially infectious materials, contaminated surfaces, or equipment. In some cases, wearing two pairs of gloves may be appropriate. Gloves should be disposed of when overtly contaminated, and removed when work with infectious materials is completed or when the integrity of the gloves is compromised. Disposable gloves should not be washed, reused or used when touching “clean” surfaces (keyboards, telephones, etc.) and they should not be worn outside the lab. Alternatives to latex gloves should be available. Hands must be washed following removal of gloves.
Mechanical pipetting devices should be available - mouth pipetting is not permitted.
An eyewash station must be readily available.
Each lab must contain a sink for hand washing.
Special care must be taken with any contaminated sharp item. Needles and syringes and other sharp items should be restricted in the laboratory for use only when there is no alternative. Plasticware should be substituted for glassware whenever possible. All sharps (e.g., syringes, needles, glass etc.) must be disposed of via the campus sharps disposal program. Refer to the Fact Sheet Handling and Disposal of Laboratory Sharps for information on the campus sharps program, available online at: http://www.drs.illinois.edu/bss/factsheets/.
Biological Waste Disposal
All non-sharps materials coming into contact with the research material must be autoclaved or disinfected prior to disposal. If bags displaying the international biohazard symbol are used to collect these materials, they must be over bagged with an opaque trash bag following autoclaving and prior to disposal in the solid waste stream. The optimal conditions for autoclaving biohazardous waste are half-loaded, single polypropylene bags, closed with elastomeric bands, without added water or incisions made in the top, free standing on a tray during processing, at 123°C for 70 minutes, excluding cooling period.
Disposal of Pathological Waste
Pathological waste includes animal carcasses, tissues and organs and human tissues and organs. University policy requires certain types of pathological waste be disposed of by incineration. All of the following animals and tissues or organs from these animals must be incinerated:
- Any animal inoculated with infectious agents
- Transgenic animals, potentially transgenic animals, “no-takes” in the production of transgenic animals, and off-spring of transgenic animals.
- All sheep and goats
- Small research animals (e.g., cats, dogs, rabbits, rats, mice, birds, etc.)
- Human tissues or organs
There are no exceptions to this policy without prior notification and approval by the Institutional Biosafety Committee.
If pick-up is needed, schedule by calling 244-7770. If more than 400 pounds of waste is going to be generated in a one week time period, the Principal Investigator must provide three weeks notice.
Materials for incineration must be packaged and labeled by the generating laboratory. Place no more than thirty pounds of pathological waste in a thick (3-4 mil) black or brown opaque bag. Twist the top closed, fold it over at the top, and tape securely with nylon filament or duct tape. Double bagging may be necessary to prevent leakage and/or protrusion of sharp edges. The bag must be labeled with the laboratory director's name, department, and the statement, "Pathological Waste for Incineration." Materials not packaged or labeled properly may be refused. Refrigerate or freeze waste between disposal and pickup.
Bedding from animals inoculated with infectious agents should be treated and disposed by incineration. Package bedding as described above. Label the bags with the laboratory director's name, department, and phone number and the statement, "Bedding for Incineration." Materials not packaged or labeled properly may be refused.
Requirements for Research Involving Human Materials
Researchers working with human materials including blood, cell lines or tissue culture media derived from human source material are subject to federal and state regulations on the Occupational Exposure to Bloodborne Pathogens. These regulations require that all employees who handle these materials be provided training and other protective measures. This type of training is mandatory for laboratories using human cell lines, human blood/plasma and other potentially infectious human materials (unfixed human tissues), or products derived from human source material. Therefore, all project personnel must take the training course entitled Safe Handling of Human Cell Lines/Materials in a Research Laboratory (formerly entitled Laboratory – Occupational Exposure to Bloodborne Pathogens). Information on training sessions is available at: http://www.drs.illinois.edu/bss/training/.
All laboratory procedures must be conducted at Biosafety Level 2 and in compliance with the University of Illinois at Urbana-Champaign Bloodborne Pathogen Exposure Control Plan (ECP). A copy of campus-wide ECP must be readily available in the lab. For additional copies, call 333-2755 or via e-mail.
As outlined in the ECP, employees (including those students who are compensated for their work in the laboratory) who handle human cell lines or unfixed human tissue must be offered the hepatitis B vaccination series. The requirement for vaccination may be waived if the Principal Investigator provides documentation that the human materials are free of the hepatitis B virus. Details on payment and scheduling of the vaccination series is provided in Chapter 6 of the ECP.
Other Biosafety Facts Sheets are available from the Biological Safety Section at our website: http://www.drs.illinois.edu/bss/factsheets/.